This human laminin 332 protein was purified from the conditioned medium of immortalized 184A1 mammary epithelial cells via affinity and size-exclusion chromatography.
Laminin 332 (formerly laminin 5) is a heterotrimeric basement membrane protein composed of the alpha3, beta3 and gamma2 chains of laminin. It is expressed in skin, breast and other tissues, where it is involved in formation of hemidesmosomes and wound healing. Laminin 332 has been implicated in cancer progression. In breast cancer, laminin 332 has been identified as a microenvironmental protein which can act as a motility factor.
From the laboratory of Philip M. Carpenter, MD, University of Southern California.
|Name:||Human Laminin 332 (Alternative names: laminin 5,laminin V, epiligrin, kalinin, ladsin, nicein).|
|Accession ID:||Alpha 3: Q16787, NP_937762.1; Beta 3: Q13751, NP_000219.2; Gamma 2: Q13753, NM_005562|
|Source:||184A1 human mammary epithelial cell conditioned media|
|Molecular Weight:||450 kDa, unprocessed|
|Amino Acid Sequence:||Sequences available upon request.|
|Purity:||Approximately 90%, Assessed by SDS PAGE.|
|Buffer:||150 mM NaCl, 10 mM Tris, pH 7.4, filter sterile|
|Stability:||Stable when kept at -20C (or lower), at the concentration provided. IMPORTANT: Rapidly lose in ability to provide cell detachment when stored after being diluted. It is suggested to thaw once into aliquots prior to dilution and diluting the aliquots immediately as needed for experiments. Aliquots should be stored in sterile polycarbonate ependorff tubes.|
|Concentration:||See product packaging. Concentration determined by OD280 of a standard curve of Bovine Seum Albumin (Concentration ≈ OD280 x 1.788)|
|Comments:||Filter Sterile, both processed and unprocessed forms are present.|
|Storage:||-20C or lower (Thawed protein retains activity for 1 month if stored at 4 C).|
Motility and invasion by purified laminin 332
A scattering assay done using purified laminin 332 (designated laminin 5 in the figure) showed dose-dependent motility.
Adapted from: Carpenter PM, Dao AV, Arain ZS, et al. Motility induction in breast carcinoma by mammary epithelial laminin 332 (laminin 5). Mol Cancer Res. 2009;7(4):462-75.
Scattering Assay: 10 ug/mL will induce scattering of 50% of MCF-7 cells after 24 hour incubation. For more details/methods on the scattering assay see: Carpenter PM, Dao AV, Arain ZS, et al. Motility induction in breast carcinoma by mammary epithelial laminin 332 (laminin 5). Mol Cancer Res. 2009;7(4):462-75.
Coating Dishes/Plates: The protein storage buffer (150 mM NaCl, 10 mM Tris, pH 7.4) will work fine, as well as PBS, hepes, or RPMI. It is suggested to start at 10 ug/ml or 100 ng in 100ul of buffer for a 96 well plate, and optimize from there. Allow 4 hours for coating, but overnight is better.
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